甘草查尔酮A对卵巢癌细胞株HO8910增殖、凋亡的影响及其机制

Effects of licochalcone A on proliferation and apoptosis of human ovarian cancer HO8910 cells

目的观察甘草查尔酮A对人卵巢癌细胞株HO8910增殖、凋亡的影响,并探讨其可能机制。方法将体外培养的HO8910细胞分为观察1、2、3组和对照组。观察1、2、3组分别加入终浓度为20、40、80μmol/L的甘草查尔酮A,对照组不加药。继续培养24、48、72 h,采用MTT法测算各组细胞增殖抑制率,采用Annexin V-FITC/PI双染和流式细胞术检测各组细胞凋亡率,采用Western blotting法检测各组半胱天冬蛋白酶激活剂(Smac)、X连锁凋亡抑制蛋白(XIAP)蛋白,采用荧光比色法检测各组半胱氨酸天冬氨酸蛋白酶3(Caspase-3)活性。结果相同培养时间情况下,随着甘草查尔酮A浓度的提升,细胞增殖抑制率、细胞凋亡率、XIAP蛋白相对表达量、Caspase-3活性逐渐升高(P均<0.05),XIAP蛋白相对表达量逐渐降低(P均<0.05)。甘草查尔酮A浓度不变的情况下,随着培养时间的延长,细胞增殖抑制率、细胞凋亡率、XIAP蛋白相对表达量、Caspase-3活性逐渐升高(P均<0.05),XIAP蛋白相对表达量逐渐降低(P均<0.05)。结论甘草查尔酮A可剂量和时间依赖的抑制HO8910细胞增殖、促进HO8910细胞凋亡,其机制可能与甘草查尔酮A下调XIAP蛋白、上调Smac蛋白并增强Caspase-3活性有关。

Objective To investigate the effects of licochalcone A on the proliferation and apoptosis of human ovarian cancer cells HO8910 and its possible mechanism. Methods The HO8910 cells cultured in vitro were divided into the observation groups 1,2,3,and the control group. Cells in the observation groups 1,2,and 3 were added with 20,40,and80 mmol/L licochalcone A,and the control group was not added any drugs. At 24,48,72 h after culture,the cell growth inhibition rate of each group was detected by MTT. The apoptosis rates of HO8910 cells were measured by Hoechst33258 and Annexin V-FITC/PI staining. The expression levels of X-linked inhibitor of apoptosis protein（ XIAP） and Smac were detected by Western blotting. The activation of Caspase-3 was detected by fluorimetry. Results At the same incubation time,the growth inhibition rate,the apoptosis rate,the expression of Smac and the activation of Caspase-3 increased gradually,but the expression of XIAP decreased gradually with the increased concentrations of licochalcone A（ all P〈0. 05）.In the constant concentration of licochalcone A,over the culture time,the growth inhibition rate,the apoptosis rate,the expression of Smac and the activation of Caspase-3 increased gradually,and the expression of XIAP decreased gradually（ all P〈0. 05）. Conclusions Licochalcone A can inhibit the proliferation of HO8910 cells and promote the apoptosis of HO8910 cells in dose-and time-dependent manner. The mechanism may be that Licochalcone A can induce the up-regulation of Smac and down-regulation of XIAP,and thereby affect activation of Caspase-3.